鼻咽癌组织中miR-634水平及其对鼻咽癌细胞生长的影响及机制研究

Effect of miR-634 level on the growth of nasopharyngeal carcinoma cells and its mechanism

目的研究鼻咽癌组织中miR-634水平及其对鼻咽癌细胞生长及MEK/ERK信号通路的影响,探讨miR-634在鼻咽癌中的作用机制。方法收集鼻咽癌组织标本60例和鼻咽部非癌组织标本60例。采用逆转录-聚合酶链反应(RT-PCR)法测定鼻咽癌组织和鼻咽癌细胞中miR-634水平。将人鼻咽癌CNE-1细胞分为空白对照组、阴性对照组和过表达miR-634组。采用MTT测定细胞增殖,采用流式细胞仪检测细胞凋亡,采用Western blotting测定鼻咽癌细胞细胞外调节蛋白激酶(ERK)、磷酸化细胞外调节蛋白激酶(p-ERK)、丝裂原活化蛋白激酶(MEK)、磷酸化丝裂原活化蛋白激酶(p-MEK)蛋白水平。结果鼻咽癌组织中miR-634水平低于癌旁组织(P<0.05)。鼻咽癌细胞中miR-634水平低于正常鼻咽部上皮细胞(P<0.05)。与空白对照组和阴性对照组比较,过表达miR-634组鼻咽癌细胞中miR-634水平和细胞凋亡率升高(P<0.05);p-ERK、p-MEK蛋白水平降低(P<0.05)。转染3 d和5 d,3组细胞OD值比较差异有统计学意义(P<0.05),与空白对照组和阴性对照组比较,过表达miR-634组细胞OD值降低(P<0.05)。结论鼻咽癌组织中miR-634水平降低,过表达miR-634可抑制鼻咽癌增殖、促进细胞凋亡,其机制可能与MEK/ERK信号通路有关。

Objective To investigate the level of miR-634 in nasopharyngeal carcinoma and its effects on the cell growth and MEK/ERK signaling pathway in nasopharyngeal carcinoma, and discuss the possible mechanism. Methods Total 60 specimens of nasopharyngeal carcinoma and 60 specimens of nasopharyngeal non-cancerous tissue were collected. The level of miR-634 in nasopharyngeal carcinoma tissues and nasopharyngeal carcinoma cells were determined by reverse transcription-polymerase chain reaction(RT-PCR). Human nasopharyngeal carcinoma CNE-1 cells were divided into blank control group, negative control group and overexpressed miR-634 group. The cell proliferation was measured by MTT. The cell apoptosis was detected by flow cytometry. The levels of extracellular regulated protein kinase(ERK), phosphorylated extracellular regulated protein kinase(p-ERK), mitogen-activated protein kinase(MEK) and phosphatiated mitogen-activated protein kinase(p-MEK) proteins in nasopharyngeal carcinoma cells were determined by Western blotting. Results The level of miR-634 in nasopharyngeal carcinoma tissues was lower than that in the adjacent tissues(P<0.05). The level of miR-634 in nasopharyngeal carcinoma cells was lower than that in normal nasopharyngeal epithelial cells(P<0.05). Compared with the blank control group and the negative control group, the level of miR-634 and the apoptosis rate in the overexpressed miR-634 group were increased(P<0.05);the levels of p-ERK, p-MEK mRNA and protein were decreased(P<0.05). After transfection for 3 d and 5 d, There was significant difference in OD values among the three groups(P<0.05), compared with the blank control group and the negative control group, the OD value of the overexpress miR-634 group was decreased(P<0.05). Conclusion The level of miR-634 is decreased in nasopharyngeal carcinoma, and overexpression of miR-634 can inhibit the proliferation of nasopharyngeal carcinoma and promote apoptosis. The mechanism may be related to MEK/ERK signaling pathway.